The ubiquitin-proteasome system in retinal health and disease

The ubiquitin–proteasome system (UPS) is the main intracellular pathway for modulated protein turnover, playing an important role in the maintenance of cellular homeostasis. It also exerts a protein quality control through degradation of oxidized, mutant, denatured, or misfolded proteins and is involved in many biological processes where protein level regulation is necessary. This system allows the cell to modulate its protein expression pattern in response to changing physiological conditions and provides a critical protective role in health and disease. Impairments of UPS function in the central nervous system (CNS) underlie an increasing number of genetic and idiopathic diseases, many of which affect the retina. Current knowledge on the UPS composition and function in this tissue, however, is scarce and dispersed. This review focuses on UPS elements reported in the retina, including ubiquitinating and deubiquitinating enzymes (DUBs), and alternative proteasome assemblies. Known and inferred roles of protein ubiquitination, and of the related, SUMO conjugation (SUMOylation) process, in normal retinal development and adult homeostasis are addressed, including modulation of the visual cycle and response to retinal stress and injury. Additionally, the relationship between UPS dysfunction and human neurodegenerative disorders affecting the retina, including Alzheimer's, Parkinson's, and Huntington's diseases, are dealt with, together with numerous instances of retina-specific illnesses with UPS involvement, such as retinitis pigmentosa, macular degenerations, glaucoma, diabetic retinopathy (DR), and aging-related impairments. This information, though still basic and limited, constitutes a suitable framework to be expanded in incoming years and should prove orientative toward future therapy design targeting sight-affecting diseases with a UPS underlying basis.

Ethnobotanic importance of plants used in pigeon-breeding in Eastern Spain

Background: The importance that birds of the Columbidae family have had throughout history is visible on the Mediterranean coast. Pigeon fancying is the art of breeding and training carrier pigeons and currently, several breeds exist. The sport of racing pigeons consists in covering a distance at maximum possible speed. However, pigeon breeding has another modality called “sport pigeon”, where several males follow a female. This study focusses on ethnobotanical knowledge of native and exotic plant species that are used for diet, breeding, stimulation, healing illnesses and staining the plumage of pigeons bred in captivity. Methods: Using semi-structured interviews, we gathered information about the different plant species traditionally used for pigeon-breeding in the region of Valencia. Background material on remedies for bird illnesses was gathered from folk botanical references, local books and journals. The plant species were collected in the study area, then identified in the laboratory using dichotomous keys and vouchered in the ABH (Herbarium of Alicante University). We used Excel ® 2003 to perform a simple statistical analysis of the data collected. Results: We collected 56 species of plants (and one variety) that included 29 botanical families. The total number of species was made up of 35 cultivated and 21 wild plants. The most common were Gramineae (14 species), Leguminosae (6 species), and Compositae (4 species). Conclusions: Pigeon breeding is an immensely popular activity in Eastern Spain, and ethnobiological knowledge about breeding pigeons and caring for them is considerable. The names and traditional uses of plants depend on their geographical location, vernacular names serve as an intangible heritage. Feeding, environmental features, and genetic makeup of individuals are relevant aspects in the maintenance of avian health.

New methodology for describing the equilibrium beach profile applied to the Valencia's beaches

Mathematical models used for the understanding of coastal seabed morphology play a key role in beach nourishment projects. These projects have become the fundamental strategy for coastal maintenance during the last few years. Accordingly, the accuracy of these models is vital to optimize the costs of coastal regeneration projects. Planning of such interventions requires methodologies that do not generate uncertainties in their interpretation. A study and comparison of mathematical simulation models of the coastline is carried out in this paper, as well as elements that are part of the model that are a source of uncertainty. The equilibrium profile (EP) and the offshore limit corresponding to the depth of closure (DoC) have been analyzed taking into account different timescale ranges. The results have thus been compared using data sets from three different periods which are identified as present, past and future. Accuracy in data collection for the beach profiles and the definition of the median grain size calculation using collected samples are the two main factors that have been taken into account in this paper. These data can generate high uncertainties and can produce a lack of accuracy in nourishment projects. Together they can generate excessive costs due to possible excess or shortage of sand used for the nourishment. The main goal of this paper is the development of a new methodology to increase the accuracy of the existing equilibrium beach profile models, providing an improvement to the inputs used in such models and in the fitting of the formulae used to obtain seabed shape. This new methodology has been applied and tested on Valencia's beaches.

Let’s get to Work! The Future of Labour in Europe. Vol. 1. CEPS Paperback, 8 September 2014

Work is both an essential part of our daily lives and one of the major policy concerns across Europe. Yet the public debate of labour issues is all too often driven by political rhetoric and short-term concerns. In this volume, researchers from seven European countries explain, in accessible language, the findings from various social sciences and what they mean for the future of labour in Europe. The conclusions they reach are addressed to policy-makers, the business world, journalists and fellow academics, and to anyone interested in the shape, size and character of the labour markets of tomorrow. “Many valuable synergies emerged between the various strands of NEUJOBS and the in-house analytical work of the European Commission.” László Andor, European Commissioner for Employment, Social Affairs and Inclusion.

Characterization of Organophosphate Pesticides in Urine and Home Environment Dust in an Agricultural Community

Thesis (Master's)--University of Washington, 2016-06

Mutational Heterogeneity in Cancer

Thesis (Ph.D.)--University of Washington, 2014-06

Validation of assembled nucleic acid-based tests in diagnostic microbiology laboratories

Medical microbiology and virology laboratories use nucleic acid tests (NAT) to detect genomic material of infectious organisms in clinical samples. Laboratories choose to perform assembled (or in-house) NAT if commercial assays are not available or if assembled NAT are more economical or accurate. One reason commercial assays are more expensive is because extensive validation is necessary before the kit is marketed, as manufacturers must accept liability for the performance of their assays, assuming their instructions are followed. On the other hand, it is a particular laboratory's responsibility to validate an assembled NAT prior to using it for testing and reporting results on human samples. There are few published guidelines for the validation of assembled NAT. One procedure that laboratories can use to establish a validation process for an assay is detailed in this document. Before validating a method, laboratories must optimise it and then document the protocol. All instruments must be calibrated and maintained throughout the testing process. The validation process involves a series of steps including: (i) testing of dilution series of positive samples to determine the limits of detection of the assay and their linearity over concentrations to be measured in quantitative NAT; (ii) establishing the day-to-day variation of the assay's performance; (iii) evaluating the sensitivity and specificity of the assay as far as practicable, along with the extent of cross-reactivity with other genomic material; and (iv) assuring the quality of assembled assays using quality control procedures that monitor the performance of reagent batches before introducing new lots of reagent for testing.

Decomposition of nitrogen-15 labeled hoop pine harvest residues in subtropical Australia

Information on decomposition of harvest residues may assist in the maintenance of soil fertility in second rotation (2R) hoop pine plantations (Araucaria cunninghamii Aiton ex A. Cunn.) of subtropical Australia. The experiment was undertaken to determine the dynamics of residue decomposition and fate of residue-derived N. We used N-15-labeled hoop pine foliage, branch, and stem material in microplots, over a 30-mo period following harvesting. We examined the decomposition of each component both singly and combined, and used C-13 cross-polarization and magic-angle spinning nuclear magnetic resonance (C-13 CPMAS NMR) to chart C transformations in decomposing foliage. Residue-derived N-15 was immobilized in the 0- to 5-cm soil layer, with approximately 40% N-15 recovery in the soil from the combined residues by the end of the 30-mo period. Total recovery of N-15 in residues and soil varied between 60 and 80% for the combined-residue microplots, with 20 to 40% of the residue N-15 apparently lost. When residues were combined within microplots the rate of foliage decomposition decreased by 30% while the rate of branch and stem decomposition increased by 50 and 40% compared with rates for these components when decomposed separately. Residue decomposition studies should include a combined-residue treatment. Based on C-15 CPMAS NMR spectra for decomposing foliage, we obtained good correlations for methoxyl C, aryl C, carbohydrate C and phenolic C with residue mass, N-15 enrichment, and total N. The ratio of carbohydrate C to methoxyl C may be useful as an indicator of harvest residue decomposition in hoop pine plantations.

Long-term evaluation of AAV-mediated sFlt-1 gene therapy for ocular neovascularization in mice and monkeys

Vascular endothelial growth factor (VEGF) is one of the major mediators of retinal ischemia-associated neovascularization. We have shown here that adeno-associated virus (AAV)-mediated expression of sFIt-1, a soluble form of the Flt-1 VEGF receptor, was maintained for up to 8 and 17 months postinjection in mice and in monkeys, respectively. The expression of sFIt-1 was associated with the long-term (8 months) regression of neovascular vessels in 85% of trVEGF029 eyes. In addition, it resulted in the maintenance of retinal morphology, as the majority of the treated trVEGF029 eyes (75%) retained high numbers of photoreceptors, and in retinal function as measured by electroretinography. AAV-mediated expression of sFIt-1 prevented the development of laser photocoagulation-incluced choroidal neovascularization in all treated monkey eyes. There were no clinically or histologically detectable signs of toxicity present in either animal model following AAV.sFlt injection. These results suggest that AAV-mediated secretion gene therapy could be considered for treatment of retinal and choroidal neovascularizations.

Increasing male involvement in family planning decision making: trial of a social-cognitive intervention in rural Vietnam

We tested a social-cognitive intervention to influence contraceptive practices among men living in rural communes in Vietnam. It was predicted that participants who received a stage-targeted program based on the Transtheoretical Model (TTM) would report positive movement in their stage of motivational readiness for their wife to use an intrauterine device (IUD) compared to those in a control condition. A quasi-experimental design was used, where the primary unit for allocation was villages. Villages were allocated randomly to a control condition or to two rounds of intervention with stage-targeted letters and interpersonal counseling. There were 651 eligible married men in the 12 villages chosen. A significant positive movement in men's stage of readiness for IUD use by their wife occurred in the intervention group, with a decrease in the proportions in the precontemplation stage from 28.6 to 20.2% and an increase in action/maintenance from 59.8 to 74.4% (P < 0.05). There were no significant changes in the control group. Compared to the control group, the intervention group showed higher pros, lower cons and higher self-efficacy for IUD use by their wife as a contraceptive method (P < 0.05). Interventions based on social-cognitive theory can increase men's involvement in IUD use in rural Vietnam and should assist in reducing future rates of unwanted pregnancy.

Cholesterol-induced caveolin targeting to lipid droplets in adipocytes: A role for caveolar endocytosis

We have investigated the targeting of caveolin to lipid bodies in adipocytes that express high levels of caveolins and contain well-developed lipid droplets. We observed that the lipid droplets isolated from adipocytes of caveolin-1 knock out mice contained dramatically reduced levels of cholesterol, indicating that caveolin is required for maintaining the cholesterol content of this organelle. Analysis of caveolin distribution by cell fractionation and fluorescent light microscopy in 3T3-L1 adipocytes indicated that addition of cholesterol rapidly stimulated translocation of caveolin to lipid droplets. The cholesterol-induced trafficking of caveolins to lipid droplets was shown to be dynamin- and protein kinase C (PKC)-dependent and modulated by src tyrosine kinase activation, suggesting a role for caveolar endocytosis in this novel trafficking pathway. Consistent with this, caveolae budding was stimulated by cholesterol addition. The present data identify lipid droplets as potential target organelles for caveolar endocytosis and demonstrate a role for caveolin-1 in the maintenance of free cholesterol levels in adipocyte lipid droplets.

Disruption of BRCA I function results in telomere lengthening and increased anaphase bridge formation in immortalized cell lines

BRCA1 is a tumor suppressor that functions in controlling cell growth and maintaining genomic stability. BRCA1 has also been implicated in telomere maintenance through its ability to regulate the transcription of hTERT, the catalytic subunit of telomerase, resulting in telomere shortening, and to colocalize with the telomere-binding protein TRF1. The high incidence of nonreciprocal translocations in tumors arising from BRCA1 mutation carriers and Brca1-null mice also raises the possibility that BRCA1 plays a role in telomere protection. To date, however, the consequences for telomere status of disrupting BRCA1 have not been reported. To examine the role of BRCA1 in telomere regulation, we have expressed a dominant-negative mutant of BRCA1 (trBRCA1), known to disrupt multiple functions of BRCA1, in telomerase-positive mammary epithelial cells (SVCT) and telomerase-negative ALT cells (GM847). In SVCT cells, expression of trBRCA1 resulted in an increased incidence of anaphase bridges and in an increase in telomere length, but no change in telomerase activity. In GM847 cells, trBRCA1 also increased anaphase bridge formation but did not induce any change in telomere length. BRCA1 colocalized with TRF2 in telomerase-positive cells and with a small subset of ALT-associated PML bodies (APBs) in ALT cells. Together, these results raise the possibility that BRCA1 could play a role in telomere protection and suggest a potential mechanism for one of the phenotypes of BRCA1 deficient cells. (c) 2005 Wiley-Liss, Inc.

The novel cytoskeleton-associated protein Neuronal protein 22: Elevated expression in the developing rat brain

Neuronal development and process targeting is mediated by proteins of the cytoskeleton. However, the signaling pathways underlying these mechanisms are complex and have not yet been fully elucidated. Neuronal protein 22 (NP22) has been identified as a cytoskeleton-associated protein. It colocalizes with microtubules and actin, the two major components of the cytoskeleton. It contains numerous signaling motifs and induces process formation in non-neuronal cells. Expression of rat NP22 (rNP22) rises incrementally at specific time points during brain development, with the greatest elevation occurring during synaptogenesis in the rat brain. its neuronal localization is primarily at the plasma membrane of the soma in the embryonic brain and progresses into homogeneous expression in the postnatal rat brain. Data suggest that NP22 may play a role in mediating the molecular events governing development of the neuronal architecture. Furthermore, its sustained expression in postnatal brain implies a function in the maintenance of neuronal morphology.

Models of psychological service provision under Australia's better outcomes in mental health care program

The Access to Allied Psychological Services component of Australia's Better Outcomes in Mental Health Care program enables eligible general practitioners to refer consumers to allied health professionals for affordable, evidence-based mental health care, via 108 projects conducted by Divisions of General Practice. The current study profiled the models of service delivery across these projects, and examined whether particular models were associated with differential levels of access to services. We found: 76% of projects were retaining their allied health professionals under contract, 28% via direct employment, and 7% some other way; Allied health professionals were providing services from GPs' rooms in 63% of projects, from their own rooms in 63%, from a third location in 42%; and The referral mechanism of choice was direct referral in 51% of projects, a voucher system in 27%, a brokerage system in 24%, and a register system in 25%. Many of these models were being used in combination. No model was predictive of differential levels of access, suggesting that the approach of adapting models to the local context is proving successful.

The information specialist in the clinical team: Building expertise in evidence-based healthcare

The Clinician Development Program (CDP) is an initiative of Queensland Health’s Quality Improvement and Enhancement Program. At the Royal Brisbane & Royal Women's Hospital Health Service Districts, evidence-base practice (EBP) is an important CDP area in which several projects were carried out in 2002. This paper describes one such project. A medical librarian was invited to accompany the clinical team on morning rounds in the Medical Assessment & Planning Unit (MAPU). The librarian conducted information skills training in the ward and helped clinicians to answer questions directly related to patient care. Questions not answered during the round were followed-up, usually within 48 hours, and responses emailed to the consultant who led the rounds. At the project’s conclusion the librarian was invited to continue as a member of the MAPU clinical team, thus acknowledging the valuable role an information specialist can play in incorporating research evidence into patient care. Clinical librarianship (CL) creates a space, albeit a contentious one, for the health librarian at the bedside. This paper describes an Australian CL project and attempts to demystify the role of an information specialist in EBP. It also highlights some of the challenges facing librarians and clinicians attempting to embed EBP in clinical settings.